Chapter IV
PRESENTATION, ANALYSIS AND INTERPRETATION OF DATA
This chapter presents the results on quantified value of beta – glucan and beta – carotene from Galiella sp. extracts.
Analysis of beta – glucan contents
Table 1. Quantified amount of beta – glucan from thallus
The table shows the mean of three trials and general mean as quantified value of beta – glucan using the microwave assisted beta – glucan extraction method. The values were labeled as g g-1(gram per 1 gram)
Basing from the result gathered, the value of quantified beta – glucan from Galiella having the mean of 0.0597g g-1(5.97%) is lesser than the amount extracted from Pleurotus ostreatus (89.2 g kg-1), Pleurotus eryngii (60 g kg -1), Pleurotus sajor-caju (70.3 g kg -1) and L. edodes (81. 2 g kg-1) as reported by Toledo et al.. (2013) using the enzymatic method and quantified using HPLC. Likewise, the beta – glucan from Galiella sp. is also lower than the quantified amount of Synytsya et al.. (2008) from P.eryngii (33.6 % to 66.4 %) and P. ostreatus (44.2 to 90.1 %). Water and alkali extraction was used but the first method was reported to have lower yield than the second in beta – glucans from P.eryngii (33.6 %). Just like in P.eryngii, P. ostreatus had the lowest yield of beta – glucan using the water extraction.
On the contrary, Bak et al.. (2014) reported a lower beta – glucan content of L. edodes (20.06 % to 44.21 %) from the mushroom’s pileus and 29.74% to 56.47 % from the stipe which is nearly 50% lower than the conclusion of Toledo et al.. (2013) of the same extraction which is the Megaenzyme kit. Also, Toledo et al.. (2013) reported that there is no significant difference between the uses of spectrophotometer versus HPLC but is not applicable in comparing the two results. The use of dry weight was accustomed in the purpose of using another formula to calculate beta –glucan.
Compared to Shiitake mushrooms with 30g (Handayani et al., 2012), Galiella sp. has higher beta – glucan. Likewise with snow mushroom and shiitake mushrooms with 0.01 % and 0.67 % (Ko & Lin, 2004). Concentration of beta – glucanfrom P. ostreatus, P. eryngii, P. pulmunaris and L. edode which contain a dry weight of 0.21 to 0.53 g per 100 g using enzymatic method by Manzi and Pizzoferrato (1999) is lower than the amount from Galiella even though enzymes were used.
The microwave assisted method to extract beta – glucan from Galiella is stressed due to the report of Villares et al.. (2012) that it can yield 100%. Also, some used alkali and acidic method but there is no jelly formation of beta – glucan from Galiella after dropped with KOH and citric acid which was done for the optimization of extracting method. When compared to other works, hot water was used which was termed as the conventional method also yield an amount of it but in lower quantities. The exposure of sample to increased high temperature helped in disrupting cell structure allowing it to be combined with water (Villares et al., 2012).
Detection of beta – carotene
Detection of the beta –carotene from the Galiella sp. heptane extract was done to prove that this terpenoid is positive. This is safe evidence when spots produced is bluish to violet in color. In the study, spots were observed when compared to Retionol Palmitate Afaxin 50,000 IU beta –carotene, though the spots where not heavy but still visible. Figures 3 and 4 show the similarities of violet spots. S stands for the standard while GE3 stands for the Galiella extract.
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| Figure 4. Positive Retinol palmitate (Standard) | Figure 5. Galiella extract with positive beta – carotene (circled area) |
Analysis of beta – carotene
Beta – carotene was quantified through spectrophotometer with the absorbance of 436 nm. The calculated values basing from the formula of Cyanotech Corporation 2002 were present in Table 2. First, percentage was calculated then converted to how many micrograms are present from 1 gram of sample. Methanolic extraction was first done (Norrizah et al., 2012; cyanotech Corporation, 2002) to eliminate other compounds. Three liquids can dissolve chloroform, acetone and hexane or any petroleum ether available (Amaya, 2001). The process was done in low light or avoiding direct light which can be a reason for the destruction of carotenoids (Cyanotech corporation, 2002; Amaya, 2001). Also, heptane and cold storage area were used to maintain beta – carotene stability (Amaya, 2001).
Table 2. Quantified beta – carotene content of Galiella sp.
The quantified amount of beta – carotene from Galiella extract is approximately 2.7591 µg / g with the absorbance of 0.005. When compared to C. cibarius containing 5.6 µg / g as reported by Kuka et al., 2014, the beta – carotene from Galiella is lower by 50%. Likewise, Robaszkiewicz et al., 2010 concluded that thirteen Polish mushrooms contain 0.233 to 18.649 µg from methanolic extract and 0.001 to 3.382 µg / g from the aqueous extract. On the list, Tricholoma equestre has the highest amount of beta – carotene (18.65 µg / g) using the methanolic extraction and Suillus bovines (3.38 µg / g) using the aqueous which are both higher than the quantified amount from Galiella. The high amount can be helped with impruties or other carotenoids not specifically beta –carotene due to the absence of any ultra violet and visible absorbing liquid as stated by Amaya (2001). As for the study, heptane was used as an absorbing liquid for the beta – carotene which is guaranteed. But, the amount of beta – carotene from Galiella is higher than L.giganteus (1.88 µg / g) and S.imbirctus (2.53 µg / g) using the acetone : hexane extraction which is a similarity to the study.
Chapter V
SUMMARY, CONCLUSION AND RECOMMENDATION
Summary
This study pivoted on the quantification of beta – glucan and beta – carotene from extracts of Galiella fruiting bodies collected at Mt. Palali. Extraction was done at Saint Mary’s University Chemistry laboratory where the methods were guided with literatures and some modification. Three replicates where done every extraction.
Beta – glucan from Galiella thallus was found out to be .0597 g g-1 (5.97%) as the mean of three trials. Microwave assisted method was the optimized extracting mode with the absence of adjusting pH.
Beta – carotene found in Galiella was found out to be 2.7591 µg / g from the three trials. The presence of the terpenoid was proved through TLC by the terpenoid test and a positive result was obtained. This can verify the calculated values shown. The spots were compared to a standard retinol.
Conclusion
The quantified amount of beta – glucan and beta – carotene from the thallus of Galiella sp. is comparable to other mushrooms presented in the literatures.
Recommendation
Along the completion of study, the following are the recommendation which can be another source of study.
- Optimization of growing method for Galiella sp to reduce risk for the naturally growing species at the collection site.
- To test the immunologic effect of beta – glucan extracted from Galiella to rats.
- Galiella polysaccharides can also be tested for prebiotic activity.
